Skip to main content

Promiscuous and lineage-specific roles of cell cycle regulators in haematopoiesis


Haematopoietic cell number is maintained by a delicate balance between cell proliferation, differentiation and death. Gene knockout studies in mice have revealed the complex roles of cyclins, CDKs, and CDK inhibitors in regulating cell proliferation and differentiation in the haematopoietic system. These studies point to families of cell cycle regulators which display both redundant and unique roles within a lineage and developmental-stage specific manner. Moreover, the promiscuity of these cell cycle regulators is critical for haematopoietic cell proliferation and differentiation. In this review, we discuss the current evidence from mouse models that the complexity and multifarious nature of the haematopoietic system is critical for its form and function.


In complex, multi-cellular organisms, the interplay between cell proliferation, differentiation, and death determines cell number, tissue architecture and function. This dynamic equilibrium is best appreciated in tissues with high volumes of cell turnover, in particular the haematopoietic system [1, 2]. The human haematopoietic system turns over approximately 1011 blood cells everyday, and the daily loss of blood cells must be balanced by an equally high rate of cell proliferation. Moreover, proliferation must be coupled with cell differentiation to continuously reconstitute and replenish all different cell lineages of the haematopoietic system. In addition to this routine turnover, the haematopoietic system exhibits remarkable flexibility at times of physiological stress. For example, red blood cell numbers increase under conditions of hypoxia, while granulocyte, macrophage and lymphocyte populations expand during infection. In contrast, the uncontrolled proliferation of haematopoietic stem cells (HSC) and progenitors is symptomatic of hyperplastic diseases and leukaemia. Thus, despite the ability of haematopoietic cells to undergo rapid proliferation and differentiation entry into the cell cycle is under stringent control during haematopoietic cell development; preventing progenitor populations becoming exhausted as a result of continuous division by restricting entry into the cell cycle. Indeed, the optimal reconstitution of haematopoiesis is achieved by quiescent cells [3], whereas cells which have already entered the latter stages of the cell cycle are ineffective at engraftment [4]. Evidence from murine and primate studies also suggests that a pool of slowly cycling HSC may also exist [57]. Ultimately, the decision to proliferate or remain in a quiescent state is central to both the maintenance and immunological function of the haematopoietic system, and is a decision largely executed by cell cycle regulation.

The cell cycle: G1/S phase transition

Cell proliferation requires entry into and successful progression through the cell division cycle [8, 9]. The mammalian cell cycle can be divided into four phases wherein S phase (DNA-synthesis) and M phase (mitosis) are separated by two intervals, G1 (gap 1-between M and S) and G2 (gap 2-between S and M). Nevertheless, most cells are in a quiescent or resting phase termed G0. The transition through G1 into S phase of the cell cycle represents a critical period within which cells become committed to cell cycle progression versus growth arrest, becoming independent of growth factor stimuli, and as such is of particular importance to the haematopoietic system.

G1/S-phase stage transition is regulated by two families of G1 cyclins, the D-type cyclins (cyclin D1, D2, and D3) and the E-type cyclins (cyclin E1 and E2) [8] and their dependent kinases (CDK4, -6 and -2). CDK4 and CDK6, bind to and are activated by the D-type cyclins, while CDK2 activity is induced by the E-type cyclins [9]. The activities of the CDKs are regulated by two families of cyclin dependent kinase inhibitors (CKIs) the Cip/Kip family and the INK4 family. The Cip/Kip family, comprised of p21Cip1, p27Kip1 and p57Kip2 modulate the activities of cyclin D-, E- and A-dependent kinases (reviewed in [8]). The INK4 family, comprised of p15INK4b, p16INK4a, p18INK4c, and p19INK4d specifically inhibit D-type cyclin associated CDK4 and CDK6 activity by formation of binary complexes which block active cyclin-CDK complex formation. The anti-proliferative activity of INK4 proteins is also in part due to competition with Cip/Kip proteins for binding to the D-type cyclin/CDK complex, releasing the Cip/Kip proteins to inhibit cyclin E-CDK2. Upon activation cyclin D-CDK4/6 and cyclin E-CDK2 kinases phosphorylate members of the retinoblastoma (Rb) protein family, pRb, p107, and p130 early and late in G1 respectively leading to the dissociation of Rb and E2F. The release of pRb-related proteins from E2F repression induces progression from G1 to S phase.

In single cell organisms, such as yeast, one cyclin dependent kinase (CDK) is enough to mediate cell cycle progression. However, concomitant with the increased levels of complexity and control required in multicellular organisms, mammalian cells have evolved families of cell cycle regulators, including multiple CDKs, cyclins, and CDK inhibitors (CKIs). The perceived rational for having multiple families of cell cycle regulators is that redundancy enables compensation where a family member becomes deregulated. However, differential expression patterns and activities of multiple family members may also allow for an increase in functional complexity, independent of their roles as guardians against pathway deregulation. Analysis of mice deficient of cell cycle regulators has provided evidence that cell cycle regulators exhibit both redundancy and specificity in vivo dependent on cell lineage and developmental timing. Moreover, studies on single as well as combined gene deletion mice have confirmed the overlapping functions of cell cycle regulators in haematopoietic cell proliferation and development, and demonstrated that the loss of function of a cell cycle regulator can be compensated by another family member (see Table 1). However, these studies have also revealed that some cell cycle regulators have unique roles, which cannot be substituted by other family members during haematopoiesis. Examples of single and double-deletion mutants generated in mice, and the cell type affected are shown in Figure 1. In the forthcoming review we summarize the evidence that D-type cyclins, CDKS and CKIs exhibit redundant, unique, and promiscuous roles in haematopoiesis.

Table 1 A summary of phenotypes observed following deletion of cell cycle regulating genes
Figure 1
figure 1

Gene deletions affecting haematopoiesis. Gene deletions are shown next to the cell type/pathway affected. The arrow indicates whether gene deletion results in an increase in cell number/activity/maturation or a decrease in cell number/activity/maturation. HSC, haematopoeitic stem cells; CMP, common myleoid progenitor; CLP, common lymphoid progenitor; MEP, megarkaryocyte/erythroid progenitor; GMP, granulocyte-macrocyte progenitor; CDK; cyclin-dependent kinase.

D-type cyclins

The D-type cyclins are the first components of the cell cycle machinery to become induced in response to mitogen stimulation, and provide a link between the extracellular mitogenic environment and the cell cycle. Two of the three D-type cyclins are usually expressed in every mammalian cell type, and the presence of multiple D-type cyclins is suggestive of functional redundancy. Indeed, the ablation of all D-type cyclins does not result in apparent defects in most other cell types and tissues until late in embryonic development (E17.5) where anaemia and cardiac defects are observed (see Table 1). Paradoxically, mouse deletion experiments have established that whilst loss of one or two of the D-type cyclins has little effect on the development of the haematopoietic system, simultaneous deletion of all D-type cyclins completely suppresses haematopoiesis, and prevents the expansion of haematopoietic stem cells [10]. These studies suggest that the inability of other cyclin-CDK kinase complexes to compensate for the loss of cyclin D activity is unique to the haematopoietic system. However, although these studies have established the overall importance of the D-type cyclin family in haematopoiesis, the specific roles of individual D-type cyclins are only beginning to emerge.

Cyclin D1

Cyclin D1 is not normally expressed in the majority of haematopoietic lineages, despite its collaborative role in haematopoietic stem cell expansion [10]. However, cyclin D1 expression has been detected in B-cell malignancies. In contrast, cyclin D2 and D3 co-express in a large number of haematopoietic cell types and are implicated in the proliferation and differentiation of most haematopoietic cells. Consistent with this hypothesis both cyclin D2 and D3 are essential for haematopoietic cell proliferation. However, cyclin D2 or D3 null mice do not demonstrate overt haematopoietic defects probably due to the fact that compensation exists between these two D-type cyclins.

Cyclin D2

Detailed analysis of different haematopoietic compartments has revealed unique tissue-specific roles for cyclin D2 in antigen-dependent B cell clonal expansion and CD5 B cell development. For example, B cells from cyclin D2-/- mice demonstrate a specific requirement for cyclin D2 in B cell receptor(BCR)-induced proliferation [11, 12]. Moreover, although conventional(B2) B cell development proceeds normally in cyclin D2 null mice, mature B cells in the spleens are hypo-responsive to BCR and mitogenic stimulation, and enter the cell cycle at a slower rate when compared with their normal counterparts in response to BCR stimulation [13]. Cyclin D2 is also important in CD5+(B1) B cell development; the CD5+ B cell compartment in the peritoneal cavity is dramatically reduced in cyclin D2 deficient mice [13]. Cyclin D2 deficient mice also exhibited fewer B cell progenitors (Sca1+B220+) but normal levels of other haematopoietic progenitor cells [14]. In addition, clonal assays demonstrated that colonies from cyclin D2 null mice were less mature (CD19lo) than those from wild-type mice (CD19hi) indicating that cyclin D2 is rate-limiting for the production of B lymphoid progenitor cells whose proliferation does not depend on BCR signaling. Normal CD5+B cells are important for eliminating bacterial pathogens through secretion of IgG3 subtype antibodies, and functioning as precursors of IgA-secreting plasma cells. Thus, cyclin D2 null mice may be expected to be deficient in host defense against bacterial pathogens; indeed, cyclin D2-/- mice display specific immunodeficiency in IgG3 as well as in IgA levels.

Cyclin D3

Cyclin D3 also has a unique role in B cell development. Analysis of cyclin D3 null mice showed that cyclin D3 was specifically required for the development of pre-B cells [15]. Consistent with this, cyclin D3, but not cyclin D2, is induced during the transition from pro-B cell to pre-B cell. Although in normal mice cyclin D2 is abundantly expressed in pro-B cells (CD43+B220+CD25-), large pre-B cells (CD43loB220+CD25lo), and immature cells (B220+IgM+) from the bone marrow, it is absent in small pre-B cells (CD43-B220+CD25+). Indeed, cyclin D3 is the only D-type cyclin expressed in these small pre-B cells, and consequently small pre-B cells are depleted in cyclin D3 deficient mice.

Cyclin D3 also has a specific role in T cell development. Cyclin D3 deficient mice display a defect in thymic T cell development characterized by a significant decrease in CD4+CD8+ double-positive (DP) T cells. This phenotype results from a non-redundant role for cyclin D3 in the maturation of CD4-CD8- double-negative (DN) T cells, which are the precursors of DP T cells. In normal mice, developing DN T cells successively pass through four stages (DN-1 to DN-4), whereas cyclin D3 deficient thymocytes fail to undergo the proliferative burst that normally occurs during the DN-3 to DN-4 transition. In addition, cyclin D3 null mice also display defects in the maturation of granulocytes in the bone marrow and have reduced levels of circulating neutrophil granulocytes [16]. As a result, cyclin D3 null mice are unable to mount a normal response to bacterial infection. Yet, the development of the myeloid progenitors proceeds relatively normally in mice lacking cyclin D3, indicating that the cyclin D3 function is only required at later stages of neutrophil development.

Compensation and redundancy of cyclin D2 and D3

Together these studies demonstrate the importance of cyclin D2 and cyclin D3 in haematopoiesis. However, some questions remain unanswered; namely, cyclin D2 is essential for B cell development, yet why do cyclin D2 null mice have a normal number of conventional (B1) B cells? In fact, induction of cyclin D3 in cyclin D2 null mice has been found to compensate for loss of cyclin D2, and allow for the development of a normal number of mature B-lymphocytes [11]. In normal mature B cells cyclin D2 is the predominant D-type cyclin and cyclin D3 is not expressed at significant levels. However, in cyclin D2 null B cells, cyclin D3 is up-regulated to compensate for the loss of cyclin D2, and has been shown to complex with CDK6 in mitogen-stimulated cells. This was the first demonstration that loss of a D-type cyclin causes specific expression and functional compensation by another member of the family in vivo, and thus provides a rationale for the presence of mature B-lymphocytes in cyclin D2-/- mice [11, 14].

The compensation between different D-cyclins was further confirmed by double D-type cyclin deletion experiments, generating mice expressing only one cyclin D. These studies revealed that in single-cyclin D embryos, the tissue-specific expression patterns of D-cyclins were lost and the remaining D-cyclin was ubiquitously expressed [10]. These double D-type cyclin deleted mice did not show additional haematopoietic phenotypes compared with the single D-type cyclin deficient mice, except that cyclin D2 and -D3 deletion is embryonic lethal at late developmental stages (~E18.5), and cyclin D2 and -D3 null embryos developed severe megaloblastic anaemia [10].

Cyclin-dependent kinases (CDKs)

CDK4 and -6 are the preferred catalytic subunits of cyclin D. Mice lacking the CDK4 gene display a dwarfism-like phenotype, infertility, and hypocellularity in many organs (see Table 1). However, no overt haematopoietic phenotype is observed in the CDK4-deficient mice, probably reflecting the compensation for loss of CDK4 by CDK6 [17]. In contrast, CDK6 null mice exhibit mild haematopoietic defects, including hypoplasia of thymuses and spleens, and depletion of megakaryocytes and erythrocytes [18]. Thus, it appears that CDK6 has a developmental and proliferative role specific for haematopoietic cells. Consistent with this notion, deletion of CDK6 results in delayed G1 progression in lymphocytes, but not in other cell types, such as mouse embryo fibroblasts.

Interestingly, mice defective for both CDK4 and CDK6 have a similar but milder phenotype compared with D-type cyclin deficient mice. CDK4 and -6 double mutant mice also display multi-lineage haematopoietic abnormalities, including a reduction in cellularity of lymphoid, myeloid and granulocyte-macrophage progenitors as well as a significant decrease in number of the more mature haematopoietic cells, such as monocytes, macrophage, lymphocytes and erythrocytes. Interestingly, further studies have revealed that D-type cyclins complex with CDK2 in the absence of CDK4 and -6, which may account for the less severe haematopoietic phenotype observed in cyclin-D null mice versus CDK4 and -6 null mice.

CDK2 is activated by E-type and A-type cyclins, and its activity is required for the transition through the G1/S and S phases. Unexpectedly, analysis of CDK2 deficient mice showed that CDK2 is dispensable for mouse development, suggesting functional compensation between different CDKs [19]. Although the CDK2-/- mice have reduced body sizes, CDK2 null mice do not display any haematopoietic specific defects. However, consistent with the compensation for CDK4 by CDK2, double deletions of CDK4 and CDK2 results in the reduced proliferation of multipotential progenitors, culminating in a decrease in cellularity in all haematopoietic subpopulations [20]. These findings indicate that CDK4, not only has overlapping roles with CDK6, but that in the absence of CDK4, cyclin D may cooperate with CDK2 to control haematopoietic progenitor proliferation. Moreover, it suggests that only regulators for early G1, but not later cell cycle phases have unique roles for haematopoietic development. Indeed, mice null for E-type cyclins demonstrate largely normal development but severely impaired endoreplication of trophoblast giant cells and megakaryocytes; thought largely to reflect a failure to incorporate MCM helicase into DNA replication origins during G1/S phase progression [21, 22]. However, whilst cyclin A1/2, cyclin B1/2/3 and cdc2(CDK1) are important cell cycle regulators for G2 and M phase progression, single gene deletion mice for these cell cycle regulators have no obvious haematopoietic phenotypes.

The Cip/Kip family of CKIs

The Cip/Kip family of proteins are potent inhibitors of CDK2 kinases and may also stabilize cyclin D-CDK4/-6 complex formation. Despite p57Kip2 being previously reported to be induced by transforming growth factor β (TGF-β) in primary human CD34+ umbilical cord blood progenitor/stem cells (CB-CD34) [23], p21Cip1 and p27Kip1 are the predominant CKIs expressed in haematopoietic cells (see Table 1).

It is generally believed that p21Cip1 and p27Kip1 set the stoichiometric thresholds for cell cycle entry and progression through G1 in response to mitogenic stimuli, such as cytokines and interactions with stromal cells and the extracellular matrix. Indeed, recent evidence suggests that p21Cip1 and p27Kip1 are critical regulators of the increased HSC-specific entry into the cell cycle observed in FoxO deficient mice [24, 25], implicating p21Cip1 and p27Kip1 in maintenance of HSC quiescence. Interestingly, phosphatase and tensin homologue (PTEN), which functions as a negative regulator of the phosphatidylinositol-3-OH kinase (PI(3)K)-Akt pathway, a known negative regulator of FOXO, may also restrict HSC activation [26].

Experimental evidence from p21Cip1 and p27Kip1 deficient mice suggests that p21Cip1 and p27Kip1 have specific roles in the regulation of quiescence of HSC and progenitors, respectively. For instance, p21Cip1 knockout mice exhibit an increase in HSC cycling and exhaustion upon transplantation, supporting a role for p21Cip1 in stem cell quiescence. Furthermore, p21Cip1 may promote DNA repair, thereby maintaining stem cell integrity. In contrast, p27Kip1 knockout mice display increased progenitor cell activity, and evidence suggests that whilst p27Kip1 controls progenitor cell replication, it does not control HSC pool size [27, 28]. Paradoxically, haematopoietic progenitor cells express p21Cip1 at a high level, and although p21Cip1 maybe sequestered from CDK2 in these cells, p21Cip1 null mice display reduced progenitor cell replication, suggesting p21Cip1 expression is required for progenitor cell replication. Interestingly, both the p21Cip1 and p27Kip1 deleted mice retain multi-lineage haematopoietic differentiation potential [29]. The mechanisms by which p21Cip1 and p27Kip1 gene inactivation differentially affects HSC and progenitors are not clearly understood. One possibility is that p21Cip1 specifically binds and regulates cellular molecules through domains that are not found in p27Kip1 and vice versa. Indeed, p21Cip1 and p27Kip1 differ substantially at their carboxy-terminus, and it is possible that domains located at this region confer specific functions [8].

Although the paradigm for Cip/Kip activity is to prevent cell cycle progression through inhibiting the CDK2 kinase activity, recent evidence demonstrated that CDK2 is dispensable for cell cycle inhibition mediated by the CKIs p27Kip1 and p21Cip1 as the loss of p27Kip1 and p21Cip1 confers similar proliferative advantages to cells with or without CDK2 [30]. These data suggests that either p27Kip1 or p21Cip1 have CDK2-independent cell cycle functions, or cells possess compensatory mechanisms that efficiently bypass their requirement for CDK2.

Aside from a decrease in HSC renewal capacity, mice lacking p21Cip1 have no significant haematopoietic defects, except that they have a substantial decrease in inflammatory circulating monocytes in peripheral blood [31]. In contrast, mice lacking p27Kip1 display multiple organ hyperplasia, with the most pronounced hyperplasia observed in the haematopoietic organs, such as thymus and spleen. Thymic hyperplasia in p27Kip1 null mice was associated with increased T lymphocyte proliferation as a result of decrease in growth factor dependency. In the spleen, the absence of p27Kip1 selectively enhanced proliferation of haematopoietic progenitor cells. Ex vivo proliferative assays also demonstrated that T and B lymphocytes lacking p27Kip1 are hypersensitive to antigen receptor and/or mitogen stimulation. Thus, p27Kip1 deficiency may cause an enhanced proliferation in response to mitogens in progenitors and more mature haematopoietic cells [3234].

The INK4 family of CKIs

The INK4 family have an essential role in G1 cell cycle phase progression through specifically inhibiting cyclin D-CDK4/6 kinase activity.

p16INK4a is highly expressed in HSC, and down-regulated with differentiation of all lineages [35]. Loss of p16INK4A in vitro increases the growth rate of myeloid colonies, and confers an increased ability for clonal expansion of hamatopoetic progenitor cells [36]. However, while it influences the growth and self-renewal kinetics of haematopoietic stem cells, p16INK4a deficient mice do not exhibit abnormal haematopoiesis [37], suggesting its role in blood cell differentiation is not essential. Mice lacking p16INK4a are not predisposed to cancer, except for a small percentage of animals that develop B-cell lymphomas after a long latency [38, 39]. Furthermore, whilst p16INK4a homozygous mutations are observed in 35% of acute lymphoblastic leukaemia (ALL) patients, p16INK4a deletion is not an independent prognostic factor in ALL [40].

In contrast, p15INK4b is absent in HSC, and its level increases in myeloid and lymphoid lineages [35, 41]. Although hypermethylation of the p15INK4b gene promoter region is a common event in acute myeloid leukaemia, transformation of myeloid cells by deregulated c-Myc is not thought to require inactivation of p15INK4b [42]. However, as p15INK4b is regulated by TGF-β1, which may maintain HSC quiescence independently of p21Cip1 or p27Kip1 [43], the role of p15INK4b in stem cell fate requires further investigation. Interestingly, p15INK4b null mice do not display gross developmental defects, except for extramedullary haematopoiesis and lymphoid hyperplasia in spleen and lymph nodes [44], suggesting that p15INK4b has specific roles in regulating the proliferation of more mature haematopoietic cells.

p18INK4c and p19INK4d are expressed at higher levels in HSC compared with more mature myeloid and lymphoid cells [45]. In addition, the G1 arrest necessary for the terminal differentiation of B lymphocytes to immunoglobulin secreting plasma cells requires p18INK4c [4547]. In contrast, mice lacking p19INK4d show normal haematopoiesis and both lymphoid and myeloid compartments differentiate normally [48]. Predictably, mice lacking p18INK4c develop widespread hyperplasia and organomegaly similar to those developed by p27Kip1 deficient mice demonstrating lymphoproliferative disorders and T cell lymphomas [44, 49]. In addition, p18INK4 deleted mice exhibit disproportionately enlarged and hyperplastic spleen and thymus. Although T and B lymphocytes develop normally in p18INK4c deficient mice, they both exhibit increased cellularity and hypersensitivity to mitogenic stimulation.

Despite p15INK4b and p18INK4c single gene deletion mice having similar haematopoietic phenotypes, combined deletion of p15INK4b and p18INK4c does not result in a more severe phenotype, suggesting that p15INK4b and p18INK4c may function in different haematopoietic cell types and have cell-type specific functions [44]. Loss of p18INK4c in T cells leads to hyperproliferation in response to T cell receptor (TCR) and/or co-receptor stimulation. These data suggest a model in which p18INK4c functions as an inhibitory threshold in quiescent T cells and modulates proliferation of T cells [50]. Interestingly, p18INK4c deleted HSC retain multilineage differentiation potential and are protected from exhaustion caused by p21Cip1 deficiency, likely by a counteracting mechanism against cellular senescence, indicating that p18INK4C has a role in restricting cell cycle entry and thus the pool size of HSC [51, 52].

Cooperation between Cip/Kip family of CKI and pRb-related proteins

Recent combined gene knock-out experiments have alluded to promiscuous roles of cell cycle regulators during haematopoiesis. The lack of overt proliferative phenotypes in the major haematopoietic organs of the p27Kip1-/-mice indicates functional redundancy and compensation exists to prevent excess uncontrolled proliferation. Indeed, analysis of p27Kip1 and p130 deficient mice have revealed that the pRb-related protein p130 can cooperate with p27Kip1 to regulate the proliferation of a number of haematopoietic lineages [53].

In p27Kip1 and p130 double deficient mice, the cellularity of the spleens but not the thymuses is significantly increased compared with their p27Kip1-/-counterparts, affecting the lymphoid, erythroid and myeloid compartments [53]. In particular, in vivo cell proliferation is significantly augmented in the B and T cells, monocytes, macrophages and erythroid progenitors suggesting that p130 can compensate in part for the loss of p27Kip1. In fact, analysis of p27Kip1 deficient splenocytes has shown that p130 can function as a CDK2 inhibitor and can compensate for the loss of p27Kip1. However, p130 and p27Kip1double deficient thymocytes still lack a significant proliferative phenotype. This observation may be in part explained by the report that p107, like p130, can also interact with CDK2 in the spleen and thymus of p27Kip1 null mice, suggesting that p107 may also be able to compensate for the absence of p27Kip1 to modulate CDK2 activity. Interestingly, in the thymus the amount of p107 binding to CDK2 increases when p27Kip1 is absent, suggesting that competitive binding of p107 and p27Kip1 to CDK2 may exist.

Thus, this compensatory mechanism can help to explain the lack of a significant proliferative phenotype in the p130 and p27Kip1double deficient thymocytes and possibly, for the absence of even more severe disorders in the splenocytes. Moreover, in the thymocytes, the expression level of p107 is high when compared with the splenocytes, thus allowing p107 to play a more prominent role than p130 in inhibiting CDK2 activity. Consequently, the lack of excessive thymocyte proliferation when both p130 and p27Kip1 are lost may represent the high levels of p107 associated with CDK2 in the thymus but not the spleen [53]. It also appears that the relative levels of p107 or p130 expression in a specific tissue type dictates which of the pRb-related proteins cooperate with p27Kip1 to regulate cell cycle entry and progression. This compensatory mechanism may provide an essential proliferation control in settings where p27Kip1 is not expressed at normal levels, as is the case in many human tumours and hyperplastic conditions.


It is now evident that the haematopoietic system displays a unique and specific dependency for cell cycle regulators, which is not unexpected, given the critical role of proliferation and differentiation in haematopoiesis. Furthermore, the cell cycle proteins involved in haematopoiesis almost exclusively regulate G1 entry and transition, probably attributable to the fact that the decision for cells to continue with cell cycle progression or to differentiate is often made in G1 phase [54]. A striking observation from mouse deletion experiments is that cell cycle regulators can exhibit specific, redundant, and compensatory activities, often, but not exclusively, linked to their temporal expression patterns during haematopoietic development (Figure 2).

Figure 2
figure 2

Examples of cell cycle regulators exhibiting specificity, compensation, and redundancy during haematopoiesis. Genes demonstrating specificity, compensation, and redundancy are shown with consequences for haematopoiesis.

Indeed, the lack of overt haematopoietic defects in the single and multiple cell cycle regulator gene deficient mice reveals that compensation and functional redundancy exist between members of individual cell cycle regulator family. In most circumstances, the masked phenotypes are results of the expression of more than one family member in a particular cell type. However, in some unique situations, the loss or deregulation of a cell cycle regulator leads to the compensatory induction of another family member, for instance, the compensation for loss of cyclin D2 by cyclin D3 in B cells described previously.

Evidence from the single and multiple gene deficient mouse studies also reveals specific non-overlapping roles for distinct cell cycle regulators such as p21Cip1, p27Kip1, and cyclin D2 during haematopoiesis. However, the majority of the present evidence has pointed to the tissue-specific expression patterns rather than specialised functions of cell cycle regulators as the basis for their cell type specific roles. For example, the specific requirement of cyclin D3 for pre-B cells is due to the fact that only cyclin D3 is expressed in these cells, and in addition, cyclin D2 can substitute for most functions of cyclin D1 in mice knock-in experiments [55]. However, in the case of p21Cip1 and p27Kip1, their ability to regulate the self-renewal capacity and pool sizes of haematopoietic stem cells and progenitors, respectively, cannot be attributable to their cell type specific expression, as both are expressed in haematopoietic stem cells [1], and may reflect differential binding-specificities [8].

Most recent studies also reveal unexpected compensatory mechanisms, where the loss of one cell cycle regulator can be compensated by members of another family. For example, p130(RB2) has been shown to be able to substitute for the absence of p27Kip1. The fact that the haematopoietic phenotype in p130RB2 and p27Kip1 double mutant mice is stronger than that in p130RB2 or p27Kip1-null mice indicates that their contribution to the phenotypes observed in p130RB2 and p27Kip1 double mutant mice is additive rather than a complete functional redundancy.

Ultimately, cell cycle regulators cooperate to set thresholds for cell cycle entry and exit in different lineages and cell types although there can be no doubt that functional redundancies between family members can protect cells from the consequence of deregulated cell cycle regulators. Redundancy may also serve to generate extra layers of control and complexity, which is required in multiorganisms. This allows specific developmental and proliferative clues to be channelled to distinct tissues and cells. In fact, the unique expression patterns and specific functions of these cell cycle regulators has provided specific functions for cell cycle regulators in haematopoiesis.

Intriguingly, most of the specific haematopoietic phenotypes only come to prominence after more detailed re-examination of the gene-deletion mice. As a consequence further evidence for specific and redundant roles of the cell cycle regulators in haematopoiesis is likely to be discovered. This information will provide crucial insights for devising new therapeutic strategies against the broad range of haematopoietic diseases.


  1. Steinman RA: Cell cycle regulators and hematopoiesis. Oncogene 2002,21(21):3403–3413. 10.1038/sj.onc.1205325

    Article  CAS  PubMed  Google Scholar 

  2. Rane SG, Reddy EP: JAKs, STATs and Src kinases in hematopoiesis. Oncogene 2002,21(21):3334–3358. 10.1038/sj.onc.1205398

    Article  CAS  PubMed  Google Scholar 

  3. Gothot A, Pyatt R, McMahel J, Rice S, Srour EF: Assessment of proliferative and colony-forming capacity after successive in vitro divisions of single human CD34+ cells initially isolated in G0. Exp Hematol 1998,26(7):562–570.

    CAS  PubMed  Google Scholar 

  4. Glimm H, Oh IH, Eaves CJ: Human hematopoietic stem cells stimulated to proliferate in vitro lose engraftment potential during their S/G(2)/M transit and do not reenter G(0). Blood 2000,96(13):4185–4193.

    CAS  PubMed  Google Scholar 

  5. Bradford GB, Williams B, Rossi R, Bertoncello I: Quiescence, cycling, and turnover in the primitive hematopoietic stem cell compartment. Exp Hematol 1997,25(5):445–453.

    CAS  PubMed  Google Scholar 

  6. MacKey MC: Cell kinetic status of haematopoietic stem cells. Cell Prolif 2001,34(2):71–83. 10.1046/j.1365-2184.2001.00195.x

    Article  CAS  PubMed  Google Scholar 

  7. Mahmud N, Devine SM, Weller KP, Parmar S, Sturgeon C, Nelson MC, Hewett T, Hoffman R: The relative quiescence of hematopoietic stem cells in nonhuman primates. Blood 2001,97(10):3061–3068. 10.1182/blood.V97.10.3061

    Article  CAS  PubMed  Google Scholar 

  8. Sherr CJ, Roberts JM: CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev 1999,13(12):1501–1512.

    Article  CAS  PubMed  Google Scholar 

  9. Sherr CJ, Roberts JM: Living with or without cyclins and cyclin-dependent kinases. Genes Dev 2004,18(22):2699–2711. 10.1101/gad.1256504

    Article  CAS  PubMed  Google Scholar 

  10. Ciemerych MA, Kenney AM, Sicinska E, Kalaszczynska I, Bronson RT, Rowitch DH, Gardner H, Sicinski P: Development of mice expressing a single D-type cyclin. Genes Dev 2002,16(24):3277–3289. 10.1101/gad.1023602

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. Lam EW, Glassford J, Banerji L, Thomas NS, Sicinski P, Klaus GG: Cyclin D3 compensates for loss of cyclin D2 in mouse B-lymphocytes activated via the antigen receptor and CD40. J Biol Chem 2000,275(5):3479–3484. 10.1074/jbc.275.5.3479

    Article  CAS  PubMed  Google Scholar 

  12. Lam EW, Glassford J, van der Sman J, Banerji L, Pizzey AR, Shaun N, Thomas B, Klaus GG: Modulation of E2F activity in primary mouse B cells following stimulation via surface IgM and CD40 receptors. Eur J Immunol 1999,29(10):3380–3389. 10.1002/(SICI)1521-4141(199910)29:10<3380::AID-IMMU3380>3.0.CO;2-C

    Article  CAS  PubMed  Google Scholar 

  13. Solvason N, Wu WW, Parry D, Mahony D, Lam EW, Glassford J, Klaus GG, Sicinski P, Weinberg R, Liu YJ, Howard M, Lees E: Cyclin D2 is essential for BCR-mediated proliferation and CD5 B cell development. Int Immunol 2000,12(5):631–638. 10.1093/intimm/12.5.631

    Article  CAS  PubMed  Google Scholar 

  14. Mohamedali A, Soeiro I, Lea NC, Glassford J, Banerji L, Mufti GJ, Lam EW, Thomas NS: Cyclin D2 controls B cell progenitor numbers. J Leukoc Biol 2003,74(6):1139–1143. 10.1189/jlb.0803363

    Article  CAS  PubMed  Google Scholar 

  15. Cooper AB, Sawai CM, Sicinska E, Powers SE, Sicinski P, Clark MR, Aifantis I: A unique function for cyclin D3 in early B cell development. Nat Immunol 2006,7(5):489–497. 10.1038/ni1324

    Article  CAS  PubMed  Google Scholar 

  16. Sicinska E, Lee YM, Gits J, Shigematsu H, Yu Q, Rebel VI, Geng Y, Marshall CJ, Akashi K, Dorfman DM, Touw IP, Sicinski P: Essential role for cyclin D3 in granulocyte colony-stimulating factor-driven expansion of neutrophil granulocytes. Mol Cell Biol 2006,26(21):8052–8060. 10.1128/MCB.00800-06

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Tsutsui T, Hesabi B, Moons DS, Pandolfi PP, Hansel KS, Koff A, Kiyokawa H: Targeted disruption of CDK4 delays cell cycle entry with enhanced p27(Kip1) activity. Mol Cell Biol 1999,19(10):7011–7019.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Malumbres M, Sotillo R, Santamaria D, Galan J, Cerezo A, Ortega S, Dubus P, Barbacid M: Mammalian cells cycle without the D-type cyclin-dependent kinases Cdk4 and Cdk6. Cell 2004,118(4):493–504. 10.1016/j.cell.2004.08.002

    Article  CAS  PubMed  Google Scholar 

  19. Berthet C, Aleem E, Coppola V, Tessarollo L, Kaldis P: Cdk2 knockout mice are viable. Curr Biol 2003,13(20):1775–1785. 10.1016/j.cub.2003.09.024

    Article  CAS  PubMed  Google Scholar 

  20. Berthet C, Klarmann KD, Hilton MB, Suh HC, Keller JR, Kiyokawa H, Kaldis P: Combined loss of Cdk2 and Cdk4 results in embryonic lethality and Rb hypophosphorylation. Dev Cell 2006,10(5):563–573. 10.1016/j.devcel.2006.03.004

    Article  CAS  PubMed  Google Scholar 

  21. Geng Y, Yu Q, Sicinska E, Das M, Schneider JE, Bhattacharya S, Rideout WM, Bronson RT, Gardner H, Sicinski P: Cyclin E ablation in the mouse. Cell 2003,114(4):431–443. 10.1016/S0092-8674(03)00645-7

    Article  CAS  PubMed  Google Scholar 

  22. Geng Y, Lee YM, Welcker M, Swanger J, Zagozdzon A, Winer JD, Roberts JM, Kaldis P, Clurman BE, Sicinski P: Kinase-independent function of cyclin E. Mol Cell 2007,25(1):127–139. 10.1016/j.molcel.2006.11.029

    Article  CAS  PubMed  Google Scholar 

  23. Scandura JM, Boccuni P, Massague J, Nimer SD: Transforming growth factor beta-induced cell cycle arrest of human hematopoietic cells requires p57KIP2 up-regulation. Proc Natl Acad Sci U S A 2004,101(42):15231–15236. 10.1073/pnas.0406771101

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Lam EW, Francis RE, Petkovic M: FOXO transcription factors: key regulators of cell fate. Biochem Soc Trans 2006,34(Pt 5):722–726.

    Article  CAS  PubMed  Google Scholar 

  25. Tothova Z, Kollipara R, Huntly BJ, Lee BH, Castrillon DH, Cullen DE, McDowell EP, Lazo-Kallanian S, Williams IR, Sears C, Armstrong SA, Passegue E, Depinho RA, Gilliland DG: FoxOs Are Critical Mediators of Hematopoietic Stem Cell Resistance to Physiologic Oxidative Stress. Cell 2007,128(2):325–339. 10.1016/j.cell.2007.01.003

    Article  CAS  PubMed  Google Scholar 

  26. Zhang J, Grindley JC, Yin T, Jayasinghe S, He XC, Ross JT, Haug JS, Rupp D, Porter-Westpfahl KS, Wiedemann LM, Wu H, Li L: PTEN maintains haematopoietic stem cells and acts in lineage choice and leukaemia prevention. Nature 2006,441(7092):518–522. 10.1038/nature04747

    Article  CAS  PubMed  Google Scholar 

  27. Cheng T, Rodrigues N, Dombkowski D, Stier S, Scadden DT: Stem cell repopulation efficiency but not pool size is governed by p27(kip1). Nat Med 2000,6(11):1235–1240. 10.1038/81335

    Article  CAS  PubMed  Google Scholar 

  28. Cheng T, Rodrigues N, Shen H, Yang Y, Dombkowski D, Sykes M, Scadden DT: Hematopoietic stem cell quiescence maintained by p21cip1/waf1. Science 2000,287(5459):1804–1808. 10.1126/science.287.5459.1804

    Article  CAS  PubMed  Google Scholar 

  29. Aleem E, Kiyokawa H, Kaldis P: Cdc2-cyclin E complexes regulate the G1/S phase transition. Nat Cell Biol 2005,7(8):831–836. 10.1038/ncb1284

    Article  CAS  PubMed  Google Scholar 

  30. Martin A, Odajima J, Hunt SL, Dubus P, Ortega S, Malumbres M, Barbacid M: Cdk2 is dispensable for cell cycle inhibition and tumor suppression mediated by p27(Kip1) and p21(Cip1). Cancer Cell 2005,7(6):591–598. 10.1016/j.ccr.2005.05.006

    Article  CAS  PubMed  Google Scholar 

  31. Scatizzi JC, Hutcheson J, Bickel E, Woods JM, Klosowska K, Moore TL, Haines GK 3rd, Perlman H: p21Cip1 is required for the development of monocytes and their response to serum transfer-induced arthritis. Am J Pathol 2006,168(5):1531–1541.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  32. Fero ML, Rivkin M, Tasch M, Porter P, Carow CE, Firpo E, Polyak K, Tsai LH, Broudy V, Perlmutter RM, Kaushansky K, Roberts JM: A syndrome of multiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27(Kip1)-deficient mice. Cell 1996,85(5):733–744. 10.1016/S0092-8674(00)81239-8

    Article  CAS  PubMed  Google Scholar 

  33. Kiyokawa H, Kineman RD, Manova-Todorova KO, Soares VC, Hoffman ES, Ono M, Khanam D, Hayday AC, Frohman LA, Koff A: Enhanced growth of mice lacking the cyclin-dependent kinase inhibitor function of p27(Kip1). Cell 1996,85(5):721–732. 10.1016/S0092-8674(00)81238-6

    Article  CAS  PubMed  Google Scholar 

  34. Nakayama K, Ishida N, Shirane M, Inomata A, Inoue T, Shishido N, Horii I, Loh DY, Nakayama K: Mice lacking p27(Kip1) display increased body size, multiple organ hyperplasia, retinal dysplasia, and pituitary tumors. Cell 1996,85(5):707–720. 10.1016/S0092-8674(00)81237-4

    Article  CAS  PubMed  Google Scholar 

  35. Furukawa Y, Kikuchi J, Nakamura M, Iwase S, Yamada H, Matsuda M: Lineage-specific regulation of cell cycle control gene expression during haematopoietic cell differentiation. Br J Haematol 2000,110(3):663–673. 10.1046/j.1365-2141.2000.02253.x

    Article  CAS  PubMed  Google Scholar 

  36. Lewis JL, Chinswangwatanakul W, Zheng B, Marley SB, Nguyen DX, Cross NC, Banerji L, Glassford J, Thomas NS, Goldman JM, Lam EW, Gordon MY: The influence of INK4 proteins on growth and self-renewal kinetics of hematopoietic progenitor cells. Blood 2001,97(9):2604–2610. 10.1182/blood.V97.9.2604

    Article  CAS  PubMed  Google Scholar 

  37. Serrano M, Lee H, Chin L, Cordon-Cardo C, Beach D, DePinho RA: Role of the INK4a locus in tumor suppression and cell mortality. Cell 1996,85(1):27–37. 10.1016/S0092-8674(00)81079-X

    Article  CAS  PubMed  Google Scholar 

  38. Sharpless NE, Bardeesy N, Lee KH, Carrasco D, Castrillon DH, Aguirre AJ, Wu EA, Horner JW, DePinho RA: Loss of p16Ink4a with retention of p19Arf predisposes mice to tumorigenesis. Nature 2001,413(6851):86–91. 10.1038/35092592

    Article  CAS  PubMed  Google Scholar 

  39. Krimpenfort P, Quon KC, Mooi WJ, Loonstra A, Berns A: Loss of p16Ink4a confers susceptibility to metastatic melanoma in mice. Nature 2001,413(6851):83–86. 10.1038/35092584

    Article  CAS  PubMed  Google Scholar 

  40. Einsiedel GH, Taube T, Hartmann R, Wellmann S, Seifert G, Henze G, Seeger K: Deletion analysis of p16(INKa) and p15(INKb) in relapsed childhood acute lymphoblastic leukemia. Blood 2002,99(12):4629–4631. 10.1182/blood.V99.12.4629

    Article  CAS  Google Scholar 

  41. Teofili L, Rutella S, Chiusolo P, La Barbera EO, Rumi C, Ranelletti FO, Maggiano N, Leone G, Larocca LM: Expression of p15INK4B in normal hematopoiesis. Exp Hematol 1998,26(12):1133–1139.

    CAS  PubMed  Google Scholar 

  42. Haviernik P, Schmidt M, Hu X, Wolff L: Consistent inactivation of p19(Arf) but not p15(Ink4b) in murine myeloid cells transformed in vivo by deregulated c-Myc. Oncogene 2003,22(11):1600–1610. 10.1038/sj.onc.1206268

    Article  CAS  PubMed  Google Scholar 

  43. Cheng T, Shen H, Rodrigues N, Stier S, Scadden DT: Transforming growth factor beta 1 mediates cell-cycle arrest of primitive hematopoietic cells independent of p21(Cip1/Waf1) or p27(Kip1). Blood 2001,98(13):3643–3649. 10.1182/blood.V98.13.3643

    Article  CAS  PubMed  Google Scholar 

  44. Latres E, Malumbres M, Sotillo R, Martin J, Ortega S, Martin-Caballero J, Flores JM, Cordon-Cardo C, Barbacid M: Limited overlapping roles of P15(INK4b) and P18(INK4c) cell cycle inhibitors in proliferation and tumorigenesis. Embo J 2000,19(13):3496–3506. 10.1093/emboj/19.13.3496

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  45. Tschan MP, Peters UR, Cajot JF, Betticher DC, Fey MF, Tobler A: The cyclin-dependent kinase inhibitors p18INK4c and p19INK4d are highly expressed in CD34+ progenitor and acute myeloid leukaemic cells but not in normal differentiated myeloid cells. Br J Haematol 1999,106(3):644–651. 10.1046/j.1365-2141.1999.01617.x

    Article  CAS  PubMed  Google Scholar 

  46. Morse L, Chen D, Franklin D, Xiong Y, Chen-Kiang S: Induction of cell cycle arrest and B cell terminal differentiation by CDK inhibitor p18(INK4c) and IL-6. Immunity 1997,6(1):47–56. 10.1016/S1074-7613(00)80241-1

    Article  CAS  PubMed  Google Scholar 

  47. Tourigny MR, Ursini-Siegel J, Lee H, Toellner KM, Cunningham AF, Franklin DS, Ely S, Chen M, Qin XF, Xiong Y, MacLennan IC, Chen-Kiang S: CDK inhibitor p18(INK4c) is required for the generation of functional plasma cells. Immunity 2002,17(2):179–189. 10.1016/S1074-7613(02)00364-3

    Article  CAS  PubMed  Google Scholar 

  48. Zindy F, van Deursen J, Grosveld G, Sherr CJ, Roussel MF: INK4d-deficient mice are fertile despite testicular atrophy. Mol Cell Biol 2000,20(1):372–378.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  49. Franklin DS, Godfrey VL, Lee H, Kovalev GI, Schoonhoven R, Chen-Kiang S, Su L, Xiong Y: CDK inhibitors p18(INK4c) and p27(Kip1) mediate two separate pathways to collaboratively suppress pituitary tumorigenesis. Genes Dev 1998,12(18):2899–2911.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  50. Kovalev GI, Franklin DS, Coffield VM, Xiong Y, Su L: An important role of CDK inhibitor p18(INK4c) in modulating antigen receptor-mediated T cell proliferation. J Immunol 2001,167(6):3285–3292.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  51. Yu H, Yuan Y, Shen H, Cheng T: Hematopoietic stem cell exhaustion impacted by p18 INK4C and p21 Cip1/Waf1 in opposite manners. Blood 2006,107(3):1200–1206. 10.1182/blood-2005-02-0685

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  52. Yuan Y, Shen H, Franklin DS, Scadden DT, Cheng T: In vivo self-renewing divisions of haematopoietic stem cells are increased in the absence of the early G1-phase inhibitor, p18INK4C. Nat Cell Biol 2004,6(5):436–442. 10.1038/ncb1126

    Article  CAS  PubMed  Google Scholar 

  53. Soeiro I, Mohamedali A, Romanska HM, Lea NC, Child ES, Glassford J, Orr SJ, Roberts C, Naresh KN, Lalani el N, Mann DJ, Watson RJ, Thomas NS, Lam EW: p27Kip1 and p130 cooperate to regulate hematopoietic cell proliferation in vivo. Mol Cell Biol 2006,26(16):6170–6184. 10.1128/MCB.02182-05

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  54. Zhu L, Skoultchi AI: Coordinating cell proliferation and differentiation. Curr Opin Genet Dev 2001,11(1):91–97. 10.1016/S0959-437X(00)00162-3

    Article  CAS  PubMed  Google Scholar 

  55. Carthon BC, Neumann CA, Das M, Pawlyk B, Li T, Geng Y, Sicinski P: Genetic replacement of cyclin D1 function in mouse development by cyclin D2. Mol Cell Biol 2005,25(3):1081–1088. 10.1128/MCB.25.3.1081-1088.2005

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references


S. Myatt is a research fellow supported by Cancer Research-UK and E. Lam's work is supported by grants from Leukaemia Research Fund and Cancer Research-UK.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Eric W-F Lam.

Additional information

Competing interests

The author(s) declare that they have no competing interests.

Authors’ original submitted files for images

Below are the links to the authors’ original submitted files for images.

Authors’ original file for figure 1

Authors’ original file for figure 2

Authors’ original file for figure 3

Rights and permissions

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reprints and Permissions

About this article

Cite this article

Myatt, S.S., Lam, E.WF. Promiscuous and lineage-specific roles of cell cycle regulators in haematopoiesis. Cell Div 2, 6 (2007).

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI:


  • Cell Cycle Regulator
  • Null Mouse
  • Deficient Mouse
  • Haematopoietic Stem Cell
  • Haematopoietic Cell